杨洋, 王子佳, 李慧玉, 张瑞萍. 白桦BpMADS12基因启动子的克隆及表达分析[J]. 西南林业大学学报, 2016, 36(1): 9-15. DOI: 10.11929/j.issn.2095-1914.2016.01.002
引用本文: 杨洋, 王子佳, 李慧玉, 张瑞萍. 白桦BpMADS12基因启动子的克隆及表达分析[J]. 西南林业大学学报, 2016, 36(1): 9-15. DOI: 10.11929/j.issn.2095-1914.2016.01.002
Yang Yang1, Wang Zijia1, Li Huiyu1,Zhang Ruiping2, . Cloning and Expression Analysis of BpMADS12 Promoter from Betula platyphylla[J]. Journal of Southwest Forestry University, 2016, 36(1): 9-15. DOI: 10.11929/j.issn.2095-1914.2016.01.002
Citation: Yang Yang1, Wang Zijia1, Li Huiyu1,Zhang Ruiping2, . Cloning and Expression Analysis of BpMADS12 Promoter from Betula platyphylla[J]. Journal of Southwest Forestry University, 2016, 36(1): 9-15. DOI: 10.11929/j.issn.2095-1914.2016.01.002

白桦BpMADS12基因启动子的克隆及表达分析

Cloning and Expression Analysis of BpMADS12 Promoter from Betula platyphylla

  • 摘要: 为探究白桦BpMADS12基因的功能,克隆了BpMADS12基因上游1750bp启动子序列,通过生物信息学对顺式作用元件进行分析,并利用农杆菌花序浸染法将其遗传转化入拟南芥,然后通过β-葡萄糖苷酸酶(GUS)组织化学染色检测BpMADS12启动子的组织表达特性及干旱胁迫应答。结果表明:BpMADS12启动子序列中含有与开花、激素及干旱响应等相关的顺式作用元件;该启动子在拟南芥中的表达模式呈现为在营养生长阶段不表达,而进入生殖生长阶段后,在根、花叶、花瓣、雄蕊、雌蕊及种子等各个组织部位中均表达;PEG胁迫后处理组拟南芥中GUS表达量低于未处理组。研究显示BpMADS12基因参与白桦的开花调节、激素应答、胁迫响应(干旱)等生物学过程,对生殖生长阶段各组织器官的发育有一定的调控作用,且负调控干旱响应途径。

     

    Abstract: To characterize the function of BpMADS12, a 1 750 bp flanking sequence upstream of the translation initiation codon was cloned, and several putative cisregulatory elements were deciphered from the promoter sequence of BpMADS12 using PLACE and PlantCARE web tools, and constructed into vector pBI101, and was transferred into Arabidopsis thaliana, then investigated the expression pattern and drought stress response via histechemical GUS staining. The results showed that flowering, multiple hormoneresponsive and droughtresponsive elements were predicted in the promoter region. Histechemical GUS staining showed that the BpMADS12 promoter expressed in all tissues and organs during reproductive phase, such as roots, floral leaves, petals, stamens, pistils, seeds, and so on, instead of vegetative phase; and the GUS expression was downregulated in transgenetic Arabidopsis thaliana after PEG stress. In conclusion, BpMADS12 participates in biological processes of flowering, hormone response, drought stress response, and regulates the development of all tissues and organs during the generative growth phase to some extent, and drought responsive pathway negatively.

     

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